Quantitative determination of 5-aminolaevulinic acid and its esters in cell lysates by HPLC-fluorescence.
The development of a reliable sensitive method for the HPLC determination of 5-aminolaevulinic acid (ALA) and ALA esters in cell lysates is described. The method relies on the quantification of a fluorescent derivative of ALA following its derivatisation with acetylacetone and formaldehyde. Following this procedure it is possible to quantify ALA in cell lysates with no need for pre-purification of the sample. The method has been validated in two ranges of concentration (0.6-65 microM, 0.1-10 microg/mL, and 30-600 microM, 5-100 microg/mL), and has also been extended and validated for the determination of ALA released from ALA prodrugs after acidic hydrolysis.
Giuntini
Francesca
F
Wolfson Laboratory of Medicinal Chemistry, Department of Pharmacy and Pharmacology, University of Bath, Bath, UK.
Bourré
Ludovic
L
Macrobert
Alexander J
AJ
Wilson
Michael
M
Eggleston
Ian M
IM
D000622:Aminolevulinic Acid / Q000032:analysis* / Q000378:metabolism; D000704:Analysis of Variance; D000818:Animals; D002460:Cell Line; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004951:Esterification; D004952:Esters / Q000032:analysis; D015603:Keratinocytes / Q000737:chemistry* / Q000378:metabolism; D016014:Linear Models; D051379:Mice; D011355:Prodrugs / Q000378:metabolism; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D004952:Esters; D011355:Prodrugs; D000622:Aminolevulinic Acid